Journal of Food Composition and Analysis 33 (2014) 5558

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Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca

Short Communication

Isolation of green coffee chlorogenic acids using activated carbon

Mirna Leonor Suarez-Quiroz a, Angelina Alonso Campos a, Gerardo Valerio Alfaro a,
Oscar Gonzalez-Ros a, Pierre Villeneuve b, Maria Cruz Figueroa-Espinoza c,*
a
Instituto Tecnologico de Veracruz, Unidad de Investigacion y Desarrollo en Alimentos, Miguel Angel de Quevedo N8 2779, 91860 Veracruz, Veracruz, Mexico
b
CIRAD, UMR 1208 IATE, 2 Place Viala, F-34060 Montpellier, France
c
Montpellier SupAgro, UMR 1208 IATE, 2 Place Viala, F-34060 Montpellier, France

A R T I C L E I N F O A B S T R A C T

Article history: Chlorogenic acids, which are interesting natural antioxidants widespread in the plant kingdom, were
Received 1 November 2012 extracted and puried from Mexican green coffee beans (Coffea arabica) using different methods. The
Received in revised form 20 September 2013 nal objective was to nd an easy way to extract high-value molecules from a complex mixture, avoiding
Accepted 5 October 2013
as much as possible the use of toxic solvents. Three extraction methods (hot water at 80 8C, aqueous
methanol 70% (v/v), and aqueous isopropanol 60% (v/v)) were tested in combination with two isolating
Keywords: methods (activated carbon, different solvents). The extracted amounts of chlorogenic acids with the six
Activated carbon
treatments (4.675.87% dry basis) presented no signicant differences. The one using hot water for
Chlorogenic acids
extraction and of activated carbon for isolation, was the simplest and the most environmentally friendly.
Green coffee
Extraction Thus it can be used as a previous step to obtain from green coffee a mixture rich in chlorogenic acids
5-O-Caffeoylquinic acid which can be further fractionated to purify a specic chlorogenic acid (i.e. in this work, 5-O-caffeoyl
Isolation quinic acid using a silica gel column). Chlorogenic acids can be used as natural antioxidants in food or
Environmentally friendly non-food products. To the best of our knowledge, activated carbon has not been used to isolate
Food analysis chlorogenic acids from green coffee.
Food composition 2013 Elsevier Inc. All rights reserved.

1. Introduction
Chlorogenic acids (CGA) (mono- and di-acyl quinic acids, with
caffeic, ferulic, and p-coumaric acids as the main acylating
residues) are natural antioxidants widespread in the plant
kingdom (Clifford, 1999), and well represented in coffee beans.
Depending on the species, green coffee beans contain some 610%
(dry basis (db)) of CGA, with levels of CGA higher in Coffea robusta
beans than in C. arabica beans (Clifford, 1999; Debry, 1993; Ky
et al., 1997). The most commonly found individual CGA is 5-O-
caffeoylquinic acid (5-CQA) (Fig. 1; IUPAC numbering (IUPAC,
1976)), often called chlorogenic acid, which is usually the only one
commercially available. According to Clifford and Jarvis (1988), a
Mexican robusta green coffee contained 5.98, 1.11, and 1.20% (db)
of caffeoyl quinic acids (CQAs), feruloyl quinic acids (FQAs), and
dicaffeoyl quinic acids (diCQAs), respectively. Ky et al. (1997)
quantied CGA on a C. liberica var dewevrei, and they observed a
maximum of 6.5% of 5- and 4-CQA from a total CQAs of 7.3% (db),
and 0.76 and 1.43% (db) of total FQAs and diCQAs, respectively.
Different methods have been used to extract and isolate CGA
from green coffee. Generally, beans are rst frozen by liquid
nitrogen to minimize CGA oxidation (Colonna, 1979) and ground.
* Corresponding author. Tel.: +33 04 99 61 28 42; fax: +33 04 99 61 30 76.
E-mail address: maria.gueroa@supagro.inra.fr (M.C. Figueroa-Espinoza).
Then most of the CGA extraction methods use polluting organic
solvents as aqueous methanol (Andrade et al., 1997; Colonna,
1979; Dibert et al., 1989; Rakotomalala, 1992), aqueous methanol
and Carrez reagents (Balyaya and Clifford, 1995; Clifford et al.,
2003; Trugo and Macrae, 1984), or aqueous 2-propanol 70% (v/v)
(Morishita et al., 1984).
Adsorption can be a more environmental friendly technique
allowing the separation of selected compounds from diluted
solutions and avoiding the use of toxic solvents. It has been largely
used for the recovery of plant phenolic compounds.
Activated carbons (AC) have been used as adsorbents to
selectively separate phenolic compounds from foods or by-
products (Soto et al., 2011). Concerning the family of cinnamic
acid derivatives, AC has been used to isolate ferulic acid from an
aqueous sugar-beet pulp enzyme hydrolyzate (Couteau and
Mathaly, 1997) and from the cooking water of maize (Creppy,
2002). To the best of our knowledge, AC has not been used to
isolate CGA from green coffee beans. Reports on the use of other
kind of absorbents to recover CGA can be found in literature, i.e.
non polar resin tested on apple juice (Kammerer et al., 2007),
hydrophobic styrene-divinylbenzene copolymer used on model
solutions (Kubo et al., 2002) or on apple pomace (Schieber et al.,
2003), and polyvinylpyrrolidone (Olsson and Samuelson, 1974).
The method proposed in this work could be an environmentally
friendly procedure to extract value-added CGA compounds from
coffee industry by-products (Murthy and Madhava Naidu, 2012),

0889-1575/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.jfca.2013.10.005
56 M.L. Suarez-Quiroz et al. / Journal of Food Composition and Analysis 33 (2014) 5558
O coffee. Only the most efcient time periods were retained for the
OH OH
7
1 O
2 6
7' 2'
3 5
4 1'
OH O 9'
8'
3'
OH 6'
4'
5'
Fig. 1. Chemical structure of 5-O-caffeoyl quinic acid.
which are considered as waste materials and are widely available
in the world. CGAs are phenolic compounds of interest as they
present several biological and functional properties: antimicrobial,
antiviral, anti-mycotoxigenic, anti-carcinogenic, antioxidant, che-
lating, and ultraviolet lter (Morishita and Ohnishi, 2001; Scholz
et al., 1994; Suarez-Quiroz et al., 2004, 2013a, 2013b). CGAs could
be then used as multifunctional natural antioxidants in food, feed,
pharmaceutical, cosmetics or nutraceutical industries. In food and
feed formulations, for example, these compounds could be used as
natural antioxidant CGA-rich extracts with preservative proper-
ties.
The aim of this work was to compare six methods of extraction
and isolation of CGA from green coffee beans, in order to nd the
simplest and the most environmentally friendly to be used as a
previous step to simplify CGA fractionation. The example of 5-CQA
purication from the CGA mixture is given. To our knowledge, this
is the rst report indicating the use of AC to isolate CGA from green
coffee grains.
2. Materials and methods
2.1. Plant material
Green coffee beans (Coffea arabica) came from the Huatusco
municipality (Veracruz, Mexico), and were harvested in 2008.
2.2. Chemicals and materials
5-CQA, acetic acid, ammonium sulphate, anhydrous sodium
sulphate, butanol, ethanol 96% (v/v), ethyl acetate, formic acid,
iodine, isopropanol, methylene chloride, phosphoric acid, toluene,
celite (95% SiO2 basis), tetramethylsilane (TMS), and activated
carbon (AC) (glassy spherical powder, 212 mm, 99.95%) were
purchased from Sigma Aldrich (Toluca, Mexico), and all solvents
were of analytical grade; HPLC grade methanol from Baeyer
(Mexico City, Mexico), aluminium thin layer chromatography
(TLC) plates silica gel 60 F254 from Merck (Estado de Mexico,
Mexico).
2.3. Sample preparation
Green coffee beans were previously frozen by liquid nitrogen in
order to minimize CGA degradation (Colonna, 1979), lyophilized,
and ground with a coffee grinder (Krups North America, Inc.,
Mexico City, Mexico) to pass a 0.5 mm sieve.
2.4. Extraction and isolation of CGA
Three extraction and two isolation methods were tested in
triplicate. Different extraction times for each method were tested
(not shown) in order to obtain a maximum of CGA from green
rest of this work.
Extraction method 1. A mixture of lyophilized green coffee
powder (100 g) and distilled water (500 mL) was magnetically
stirred for 30 min at 80 8C, in the dark. After cooling, the solution
was vacuum ltered through celite (1 cm).
OH
Extraction method 2. A mixture of lyophilized green coffee
powder (100 g) and aqueous methanol 70% (v/v) (500 mL) was
magnetically stirred for 24 h at room temperature, in the dark. The
solution was then vacuum ltered through celite (1 cm), and
OH methanol was evaporated in a rotary evaporator (Bioblock, Mexico
City, Mexico).
Extraction method 3. A mixture of lyophilized green coffee
powder (100 g) and aqueous isopropanol 60% (v/v) (500 mL) was
magnetically stirred for 48 h at room temperature, in the dark. The
solution was then vacuum ltered through celite (1 cm), and
isopropanol evaporated in a rotary evaporator.
Isolation method 1 (Rakotomalala, 1992). To aqueous extract
obtained from each of the three extraction methods was added
ammonium sulphate to a nal concentration of 20 g/L, in order to
precipitate proteins by increasing the ionic strength. To make CGA
more soluble in ethyl acetate, 4% phosphoric acid was added.
Extracts were then treated three times with methylene chloride
(300 mL) to eliminate caffeine in the organic phase. The aqueous
phase containing CGA was extracted 4 times using ethyl acetate
(300 mL). The four ethyl acetate phases were pooled, dried with
anhydrous sodium sulphate (10 g), ltered (N81 lter paper,
Whatman, Mexico City, Mexico), and dried at 40 8C and 120 rpm in
a rotary evaporator. The residue was analyzed by TLC and by HPLC.
Isolation method 2 (AC). The aqueous extract obtained from
each of the three extraction methods was adjusted to pH 3.0 with
phosphoric acid, to which AC at 40 g/L was added, and
magnetically stirred for 30 min at 60 8C, under dark. After cooling
at room temperature, the mix was vacuum ltered through celite
(1 cm). CGA were desorbed from AC using ethanol 96% (v/v), then
dried with anhydrous sodium sulphate, and nally dried in a rotary
evaporator at 60 8C and 120 rpm. The residue was analyzed by TLC
and by HPLC.
2.5. Purication of 5-CQA
The 5-CQA (Fig. 1) was puried from each of the six CGA
mixtures with a silica gel 60 column (25 cm long, 1.6 cm diameter)
using toluene/ethyl acetate (90:10, v/v) as the eluted solvent.
Collected fractions were dried in a rotary evaporator at 60 8C and
120 rpm, followed by a TLC, HPLC, and NMR analysis.
2.6. Thin-layer chromatography (TLC)
TLC was performed using aluminium silica gel 60 F254 plates
(4 cm  6.6 cm). Five microliters of each sample or commercial 5-
CQA (used as the control) diluted in methanol, were spotted at
0.6 cm from the bottom of the plate. The TLC plate was then placed
in a developing chamber containing butanol/water/acetic acid
(6:2:2, v/v/v) at room temperature (Lewis et al., 1998; Lu et al.,
2004). When the solvent reached 0.4 cm from the top of the plate,
the plate was removed, dried for 5 s with a hair-dryer, observed at
254 and 360 nm, and nally revealed using iodine. The retention
factor (Rf) was calculated as the distance travelled by the individual
compound, divided by the distance travelled by the solvent. Assays
were performed in duplicate.
2.7. HPLC analysis
Diluted samples were ltered (Millex-HV, Millipore Co.)
(0.45 mm) and injected (10 mL) into an HPLC (Hypersil C18;
 M.L. Suarez-Quiroz et al. / Journal of Food Composition and Analysis 33 (2014) 5558 57

5 mm; 250 mm  4.6 mm; detection by UV absorbance at 276 nm Table 1

Green coffee chlorogenic acids extraction and isolation yields.
and room temperature). Elution was performed during 14 min
using aqueous methanol (70%, v/v) at 1 mL/min, and 40 8C (Yuan Treatment (T) Extraction/isolation method Total CGAa
et al., 2006). CGA are expressed as equivalents of 5-CQA. All (% dbb)
samples were injected in triplicate. T1 Water 80 8C/Rakotomalala (1992) 5.62  0.8c
T2 Water 80 8C/activated carbon 5.07  1.0
2.8. 1H NMR and 13
C NMR T3 Methanol 70% (v/v)/Rakotomalala (1992) 5.87  1.1
T4 Methanol 70% (v/v)/activated carbon 4.67  1.6
T5 Isopropanol 60% (v/v)/Rakotomalala (1992) 5.87  1.0
30 mg of the sample were analyzed by 1H and 13C NMR spectra T6 Isopropanol 60% (v/v)/activated carbon 5.28  1.2
were recorded using a Varian 500 MHz NMR spectrometer in a
Chlorogenic acids.
methanol-D4 solutions (Budryn et al., 2009; Hernandez et al., b
Dry basis.
2009). Chemical shifts were read relative to the internal standard c
Mean of triplicates (mean values  standard deviations).
(TMS).

3. Results and discussion Table 2

13
C NMR and 1H NMR (500 MHz, methanol-D4) chemical shifts for 5-O-caffeoyl
quinic acid.
Methods using chemical reagents or organic solvents to extract
and isolate phenolic compounds from plant extracts are not always 5-CQAa number of carbon (Fig. 1) 13
C (d/ppm) 1
H (d/ppm)
ecologically friendly. In this research, AC was tested to isolate CGA 1 74.71
from green arabica coffee beans as an alternative to limit the use of 2 37.33 2.02 br tb
polluting solvents. Three extraction methods (water at 80 8C, 2.22 br db
3 70.51 5.31 tb
aqueous methanol 70% (v/v) and aqueous isopropanol 60% (v/v))
4 72.06 3.71 db
and two isolation methods (Rakotomalala (1992) and AC) were 5 70.57 4.15 br sb
tested. Results obtained from the six extraction-isolation treat- 6 36.76 2.05 br d
ments are presented in Table 1. No signicant differences 2.14 d
7 175.61
(a = 0.05) were observed between results and the CGA content
10 126.36
varied from 4.67 to 5.87% (db). 20 113.86 7.027 br s
5-CQA was then puried in a silica gel column from the CGA 30 145.33
mixtures. The monitoring of 5-CQA purication (97% of purity) was 40 148.11
made by TLC (Rf = 0.6) and HPLC, and its structure was conrmed 50 115.07 6.94 d
60 121.58 6.75 d
by 1H and 13C NMR spectroscopy (Table 2). The puried 5-CQA was
70 145.66 7.56 d
identied by comparison with spectroscopic data reported by 80 113.79 6.22 d
Budryn et al. (2009), Hernandez et al. (2009), and Bhatt (2011). 90 167.27
The 13C NMR spectrum showed the presence of sixteen carbon a
5-O-caffeoyl quinic acid.
atoms, including two carbonyl groups at d 175.61 and d 167.27, b
br t broad triplet, br d broad doublet, t triplet, d doublet, br s broad singlet.
corresponding to carbons 7 and 90 , respectively; two aromatic
carbons bonded to hydroxyl groups at d 148.11 and d 145.33
identied as C40 and C30 ; two olenic carbons at d 145.66 and d Results are consistent with the CGA coffee content reported in the
113.79 corresponding to C70 and C80 ; four aromatic carbons literature (Clifford and Jarvis, 1988; Ky et al., 1997). Differences in
assigned to C10 , C20 , C50 , and C60 at d 126.36, d 113.86, d 115.07, and results can be explained by the small variations between methods
d 121.58, respectively; three carbons bonded to hydroxyl groups at and by the coffee variety.
d 74.71, d 70.51, and d 72.06 identied as C1, C3, and C4; one carbon The use of water at 80 8C combined with AC (treatment T2 in
bonded to an ester group at d 70.57 attributed to C5; and two Table 2) is less polluting than treatments T1 and T3 to T6, gave a
methylene identied as C2 and C6 at d 37.33 and d 36.76, good extraction yield (5.07  1.0% db), and presented no signicant
respectively. differences to alcohol extraction methods combined with AC.
The 1H NMR spectrum displayed two ortho-coupled doublet
each for 1H, at d 6.94 and d 6.75, and a broad singlet for 1H at d
7.027, conrming the presence of a tri-substituted aromatic ring; 4. Conclusion
and two doublets, each for 1H, at d 7.56 (H-70 ) and 6.22 (H-80 ),
indicating the presence of trans-di-substituted ethylene moiety in In conclusion, the use of AC to isolate CGA presented no
the molecule. The main spectroscopic data are resumed in Table 2. signicant differences with the Rakotomalala (1992) method.
Ky et al. (1997) compared different isolation methods on Adsorption with AC proved to be a suitable means to trap CGA
coffee beans previously crushed and frozen in liquid nitrogen. from a complex medium like a green coffee extract obtained either
After an aqueous methanol 70% (v/v) extraction at 4 8C overnight, by alcohol of by hot water extraction. Adsorption of CGA on AC
different methods were tested, based either on the aqueous avoids the use of toxic solvents and is a simple and less time-
extract (after evaporation of methanol): a solvent extraction with consuming method, and could be used to replace methods using
organic solvents (Rakotomalala, 1992); or a ltration through a solvents. This method is a good alternative prior to the
C18 cartridge (Bicchi et al., 1995); or a treatment by Carrez purication of 5-CQA or another single (monomer or dimer)
reagents (Balyaya and Clifford, 1995); or on the methanol CGA, which are not always commercially available, or are very
extracts (without evaporation of methanol): Carrez reagents expensive.
(Trugo and Macrae, 1984); or directly analyzed by HPLC (DIN-
10767, 1992). When using an aqueous methanol 70% (v/v) Acknowledgments
extraction combined with the Rakotomalala (1992) isolation
method, they obtained 4.1% (db) of total CGA. In our study, We thank Dr. Jorge Suarez Medelln from the University of
treatment T3 (Table 2) is similar to that used by these authors and Veracruz (Mexico) for his analytical advices. Angelina Alonso
very close results were obtained (5.87  1.1% db of total CGA). Campos thanks CONACYT for her Master degree scholarship.
58 M.L. Suarez-Quiroz et al. / Journal of Food Composition and Analysis 33 (2014) 5558
References
Andrade, P.B., Leitao, R., Seabra, R.M., Oliveira, M.B., Ferreira, M.A., 1997. Develop-
ment of an HPLC/diode-array detector method for simultaneous determination
of seven hydroxycinnamic acids in green coffee. Journal of Liquid Chromatog-
raphy and Related Technologies 20 (13) 20232030.
Balyaya, K.J., Clifford, M.N., 1995. Individual chlorogenic acids and caffeine contents
in commercial grades of wet and dry processed Indian green Robusta coffee.
Journal of Food Science and Technology-Mysore 32 (2) 104108.
Bhatt, B., 2011. Chemical constituents of Solanum xanthocarpum. Journal of Chemi-
cal and Pharmaceutical Research 3 (3) 176181.
Bicchi, C.P., Binello, A.E., Pellegrino, G.M., Vanni, A.C., 1995. Characterization of
green and roasted coffees through the chlorogenic acid fraction by HPLC-UV and
principal component analysis. Journal of Agricultural and Food Chemistry 43 (6)
15491555.
Budryn, G., Nebesny, E., Podsedek, A., Zyzelewicz, D., Materska, M., Jankowski, S.,
Janda, B., 2009. Effect of different extraction methods on the recovery of
chlorogenic acids, caffeine and Maillard reaction products in coffee beans.
European Food Research and Technology 228 (6) 913922.
Clifford, M.N., 1999. Chlorogenic acids and other cinnamates nature, occurrence
and dietary burden. Journal of the Science of Food and Agriculture 79 (3) 362
372.
Clifford, M.N., Jarvis, T., 1988. The chlorogenic acids content of green robusta coffee
beans as a possible index of geographic origin. Food Chemistry 29 (4) 291298.
Clifford, M.N., Johnston, K.L., Knight, S., Kuhnert, N., 2003. Hierarchical scheme for
LC-MSn identication of chlorogenic acids. Journal of Agricultural and Food
Chemistry 51 (10) 29002911.
Colonna, J.P., 1979. Lacide chlorogenique et les depsides de divers cafeiers africains
et malgaches: leur participation au metabolisme et leur signication biologi-
que. O.R.S.T.O.M., Paris, France.
Couteau, D., Mathaly, P., 1997. Purication of ferulic acid by adsorption after
enzymic release from a sugar-beet pulp extract. Industrial Crops and Products
6, 237252.
Creppy, E.E., 2002. Update of survey, regulation and toxic effects of mycotoxins in
Europe. Toxicology Letters 127 (1-3) 1928.
Debry, G., 1993. Le cafe et la sante..
Dibert, K., Cros, E., Andrieu, J., 1989. Solvent extraction of oil and chlorogenic acid
from green coffee. Part I. Equilibrium data. Journal of Food Engineering 10 (1)
111.
DIN-10767, 1992. Analysis of coffee and coffee products determination of
chlorogenic acids content HPLC method..
Hernandez, C.E., Chen, H.H., Chang, C.I., Huang, T.C., 2009. Direct lipase-catalyzed
lipophilization of chlorogenic acid from coffee pulp in supercritical carbon
dioxide. Industrial Crops and Products 30 (3) 359365.
IUPAC, 1976. Nomenclature of cyclitols. Biochemical Journal 153, 2331.
Kammerer, D.R., Saleh, Z.S., Carle, R., Stanley, R.A., 2007. Adsorptive recovery of
phenolic compounds from apple juice. European Food Research and Technology
224 (5) 605613.
Kubo, I., Fujita, K., Nihei, K., 2002. Anti-Salmonella activity of alkyl gallates. Journal
of Agricultural and Food Chemistry 50 (23) 66926696.
Ky, C.-L., Noirot, M., Hamon, S., 1997. Comparison of ve purication methods for
chlorogenic acids in green coffee beans (Coffea sp.). Journal of Agricultural and
Food Chemistry 45 (3) 786790.
Lewis, C.E., Walker, J.R.L., Lancaster, J.E., Sutton, K.H., 1998. Determination of
anthocyanins, avonoids and phenolic acids in potatoes. I: Coloured cultivars
of Solanum tuberosum L. Journal of the Science of Food and Agriculture 77 (1)
4557.
Lu, H.-T., Jiang, Y., Chen, F., 2004. Application of preparative high-speed counter-
current chromatography for separation of chlorogenic acid from Flos lonicerae.
Journal of Chromatography A 1026 (12) 185190.
Morishita, H., Iwahashi, H., Osaka, N., Kido, R., 1984. Chromatographic separation
and identication of naturally occurring chlorogenic acids by 1H nuclear
magnetic resonance spectroscopy and mass spectrometry. Journal of Chroma-
tography A 315 (0) 253260.
Morishita, H., Ohnishi, M., 2001. Absorption, metabolism and biological activities of
chlorogenic acids and related compounds. In: Atta ur, R. (Ed.), Stud. Nat. Prod.
Chem. Elsevier, pp. 919953.
Murthy, P.S., Madhava Naidu, M., 2012. Sustainable management of coffee industry
by-products and value additiona review. Resources, Conservation and Recy-
cling 66 (0) 4558.
Olsson, L., Samuelson, O., 1974. Chromatography of aromatic acids and aldehydes
and phenols on cross-linked polyvinylpyrrolidone. Journal of Chromatography
A 93 (1) 189199.
Rakotomalala, J.J.R., 1992. Diversite biochimique des cafeiers: analyse des acides
hydroxycinnamiques, bases puriques et diterpe`nes glycosides. Particularites
des cafeiers sauvages de la region malgache (Mascarocoffea chev.) Universite
Sciences et Techniques du Languedoc, Montpellier II, France, Montpellier,
France, pp. 219.
Schieber, A., Hilt, P., Streker, P., Endre, H.-U., Rentschler, C., Carle, R., 2003. A new
process for the combined recovery of pectin and phenolic compounds from
apple pomace. Innovative Food Science and Emerging Technologies 4 (1) 99
107.
Scholz, E., Heinrich, M., Hunkler, D., 1994. Caffeoylquinic acids and some biological-
activities of Pluchea-Symphytifolia. Planta Medica 60 (4) 360364.
Soto, M.L., Moure, A., Dominguez, H., Parajo, J.C., 2011. Recovery, concentration
and purication of phenolic compounds by adsorption: a review. Journal of
Food Engineering 105 (1) 127.
Suarez-Quiroz, M.L., Alonso Campos, A., Valerio Alfaro, G., Gonzalez-Ros, O.,
Villeneuve, P., Figueroa-Espinoza, M.C., 2013a. Anti-Aspergillus activity of green
coffee 5-O-caffeoyl quinic acid and its alkyl esters. Microbial Pathogenesis 61
62, 5156.
Suarez-Quiroz, M.L., Gonzalez-Ros, O., Barel, M., Guyot, B., Schorr-Galindo, S.,
Guiraud, J.P., 2004. Effect of chemical and environmental factors on Aspergillus
ochraceus growth and toxigenesis in green coffee. Food Microbiology 21 (6)
629634.
Suarez-Quiroz, M.L., Taillefer, W., Lopez Mendez, E.M., Gonzalez-Ros, O., Ville-
neuve, P., Figueroa-Espinoza, M.C., 2013b. Antibacterial activity and antifungal
and anti-mycotoxigenic activities against Aspergillus avus and A. ochraceus of
green coffee chlorogenic acids and dodecyl chlorogenates. Journal of Food
Safety 33 (3) 360368.
Trugo, L.C., Macrae, R., 1984. Chlorogenic acid composition of instant coffees.
Analyst 109, 263266.
Yuan, B., Qiao, M., Xu, H., Wang, L., Li, F., 2006. Determination of chlorogenic acid in
rat plasma by high performance chromatography after peritoneal administra-
tion of compound Daqingye injection. Yakugaku Zasshi Journal of the Pharma-
ceutical Society of Japan 126 (9) 811814.